Perspectives on uterine transplantation.

Uterine transplantation is a groundbreaking solution for absolute uterine factor infertility (AUFI), offering hope for motherhood. Advances in transplant surgeries and immunosuppressants have enabled thousands of women to safely become mothers. Approximately 1.5 million women globally suffer from AUFI and thus Uterine transplantation is a significant breakthrough in this arena. So far, 23 babies have been born from transplanted wombs. Uterine transplantation redefines possibilities for motherhood, empowering women's reproductive rights. Ethical considerations persist, but it marks significant progress in reproductive medicine and transplantation science. Further research is needed to evaluate risks and benefits for the future.

Genome-wide analysis of glutamate receptor gene family in allopolyploid Brassica napus and its diploid progenitors.

Ionotropic glutamate receptors are ligand-gated nonselective cation channels that mediate neurotransmission in the central nervous system of animals. Plants possess homologous proteins called glutamate receptor-like channels (GLRs) which are involved in vital physiological processes including seed germination, long-distance signaling, chemotaxis, Ca2+ signaling etc. Till now, a comprehensive genome-wide analysis of the GLR gene family members in different economically important species of Brassica is missing. Considering the origin of allotetraploid Brassica napus from the hybridization between the diploid Brassica oleracea and Brassica rapa, we have identified 11, 27 and 65 GLR genes in B. oleracea, B. rapa and B. napus, respectively showing an expansion of this gene family in B. napus. Chromosomal locations revealed several tandemly duplicated GLR genes in all the three species. Moreover, the gene family expanded in B. napus after allopolyploidization. The phylogenetic analysis showed that the 103 GLRs are classified into three main groups. The exon-intron structures of these genes are not very conserved and showed wide variation in intron numbers. However, protein sequences are much conserved as shown by the presence of ten short amino acid sequence motifs. Predicted cis-acting elements in 1 kb promoters of GLR genes are mainly involved in light, stress and hormone responses. RNA-seq analysis showed that in B. oleracea and B. rapa, some GLRs are more tissue specific than others. In B. napus, some GLRs are downregulated under cold stress, while others are upregulated. In summary, this bioinformatic study of the GLR gene family of the three Brassica species provides evidence for the expansion of this gene family in B. napus and also provided useful information for in-depth studies of their biological functions in Brassica.

ProLung™-budesonide Inhibits SARS-CoV-2 Replication and Reduces Lung Inflammation.

BACKGROUND: Inhaled budesonide benefits patients with COVID-19. ProLung™-budesonide enables the sustained, low dose administration of budesonide within a delivery vehicle similar to lung surfactant. ProLung™-budesonide may offer anti-inflammatory and protective effects to the lung in COVID-19, yet it's effect on SARS-CoV-2 replication is unknown. OBJECTIVE: To determine the efficacy of ProLung™-budesonide against SARS-CoV-2-infection in vitro, evaluate its ability to decrease inflammation, and airway hyperresponsiveness in an animal model of lung inflammation. METHODS: SARS-CoV-2-infected Vero 76 cells were treated with ProLung™-budesonide ([0.03-100 µg/ml]) for 3 days, and virus yield in the supernatant was measured. Ovalbumin-sensitized C57BL/6 mice received aerosolized (a) ProLung™-budesonide weekly, (b) only budesonide, either daily or weekly, or (c) weekly empty ProLung™ carrier (without budesonide). All treatment groups were compared to sensitized untreated, or normal mice using histopathologic examination, electron microscopy (EM), airway hyperresponsiveness (AHR) to Methacholine (Mch) challenge, and eosinophil peroxidase activity (EPO) measurements in bronchioalveolar lavage (BAL). RESULTS: ProLung™-budesonide showed significant inhibition of viral replication of SARS-CoV-2-infected cells with the selectivity index (SI) value >24. Weekly ProLung™-budesonide and daily budesonide therapy significantly decreased lung inflammation and EPO in BAL. ProLung™-budesonide localized in type II pneumocytes, and was the only group to significantly decrease AHR, and EPO in BAL with Mch challenge. CONCLUSIONS: ProLung™-budesonide significantly inhibited viral replication in SARS-CoV-2-infected cells. It localized into type II pneumocytes, decreased lung inflammation, AHR and EPO activity with Mch challenge. This novel drug formulation may offer a potential inhalational treatment for COVID-19.

Environmental externalities of the COVID-19 lockdown: Insights for sustainability planning in the Anthropocene.

The COVID-19 pandemic has abruptly halted the Anthropocene's ever-expanding reign for the time being. The resulting global human confinement, dubbed as the Anthropause, has created an unprecedented opportunity for us to evaluate the environmental consequences of large-scale changes in anthropogenic activities. Based on a methodical and in-depth review of related literature, this study critically evaluates the positive and negative externalities of COVID-19 induced lockdown on environmental components including air, water, noise, waste, forest, wildlife, and biodiversity. Among adverse impacts of the lockdown, increased amount of healthcare waste (300-400%), increased level of atmospheric ozone (30-300%), elevated levels of illicit felling in forests and wildlife poaching were prominent. Compared to the negative impacts, significant positive changes in various quality parameters related to key environmental components were evident. Positive impacts on air quality, water quality, noise level, waste generation, and wildlife were apparent in varying degrees as evaluated in this study. By presenting a critical overview of the recommendations given in the major literature in light of these documented impacts, this paper alludes to potential policy reforms as a guideline for future sustainable environmental management planning. Some of the key recommendations are e.g., enhance remote working facilities, cleaner design, use of internet of things, automation, systematic lockdown, and inclusion of hazardous waste management in disaster planning. The summarized lessons of this review, pertinent to the dynamic relationship between anthropogenic activities and environmental degradation, amply bring home the need for policy reforms and prioritization of Sustainable Development Goals in the context of the planetary boundaries to the environmental sustainability for a new post-pandemic world.

Urban metabolism of phosphorus in the food production-consumption system of Bangladesh.

Anthropogenic disruption of the global phosphorus (P) cycle has already pushed it beyond the planetary boundary. Understanding P metabolism at global, regional and local scales is critical to close the loop of P for the safekeeping of mankind. Investigating the effects of urbanization-induced income growth on the natural nutrient (especially P) cycles contribute to that end. Bangladesh, a lower-middle-income agrarian economy seeing rapid urbanization and stunning GDP growth, presents itself as a good case for P-metabolism research. Past efforts to quantify P flows in the country have not addressed the effects of urbanization thereon. This time-series study quantifies the P flows in rural and urban Bangladesh using substance flow analysis after outlining the urbanization indicators (viz. GDP, income per capita, percentage of income spent on food, change in urban population and built-up area) which affects urban metabolism of P. Urbanization caused a dietary transition from cereal-based to animal-based diet resulting in 50% more P consumption from the latter by urban individuals than their rural counterparts in 2010. Comparing the P flows among the 19 expenditure groups of the urban population, an individual belonging to a higher expenditure group (USD 71-82) consumed 38% more P than one of the lower groups (USD 17-21) in 2016. Future forecasting was conducted for (i) future demand of P fertilizer using human appropriation of net primary productivity (HANPP) and (ii) P recovery potential from urban household food waste for the policymakers to get a glimpse of the future demand and recovery potential of P. The projections suggested approximately 145% rise in the national P inflow by 2030. Moreover, the universal adoption of source separation of household food waste in the two largest cities of Bangladesh can cycle back almost 1.2 × 103 tonnes of P to the system by 2030. As Bangladesh poises to faster economic growth in decades ahead, the study provides a basis for policy formulation for an appropriate P management plan to achieve circularity in nutrient use.

Evaluation of indigenous aromatic rice cultivars from sub-Himalayan Terai region of India for nutritional attributes and blast resistance.

Indigenous folk rice cultivars often possess remarkable but unrevealed potential in terms of nutritional attributes and biotic stress tolerance. The unique cooking qualities and blissful aroma of many of these landraces make it an attractive low-cost alternative to high priced Basmati rice. Sub-Himalayan Terai region is bestowed with great agrobiodiversity in traditional heirloom rice cultivars. In the present study, ninety-nine folk rice cultivars from these regions were collected, purified and characterized for morphological and yield traits. Based on traditional importance and presence of aroma, thirty-five genotypes were selected and analyzed for genetic diversity using micro-satellite marker system. The genotypes were found to be genetically distinct and of high nutritive value. The resistant starch content, amylose content, glycemic index and antioxidant potential of these genotypes represented wide variability and 'Kataribhog', 'Sadanunia', 'Chakhao' etc. were identified as promising genotypes in terms of different nutritional attributes. These cultivars were screened further for resistance against blast disease in field trials and cultivars like 'Sadanunia', 'T4M-3-5', 'Chakhao Sampark' were found to be highly resistant to the blast disease whereas 'Kalonunia', 'Gobindabhog', 'Konkanijoha' were found to be highly susceptible. Principal Component analysis divided the genotypes in distinct groups for nutritional potential and blast tolerance. The resistant and susceptible genotypes were screened for the presence of the blast resistant pi genes and association analysis was performed with disease tolerance. Finally, a logistic model based on phenotypic traits for prediction of the blast susceptibility of the genotypes is proposed with more than 80% accuracy.

Computational analysis of the glutamate receptor gene family of Arabidopsis thaliana.

Ionotropic glutamate receptors (iGluRs) function as glutamate-activated ion channels in rapid synaptic transmission in animals. Arabidopsis thaliana possess 20 glutamate receptor-like genes (AtGLRs) in its genome which are involved in many functions including light signal transduction and calcium homeostasis. However, little is known about the physico-chemical, functional and structural properties of AtGLRs. In this study, glutamate receptor-like genes of A. thaliana have been studied in silico. Exon-intron structures revealed common origin of majority of these genes. The presence of several phosphorylation and myristoilation sites indicate the involvement of AtGLRs in various signaling processes. Gene ontology analysis showed the participation of AtGLRs in various biological processes including different stress responses. In two genes namely AT2G17260 and AT4G35290, presence of RAV1-A binding site motif in the promoter coupled with results from gene ontology annotation indicate their role in stomatal movement through abscisic acid signaling. Expression analysis showed differential expression of several tandemly arranged genes which indicates neo or sub-functionalization. Two genes namely AT5G48400 and AT5G48410 showed significantly more expression in response to Botrytis cinerea infection. Five of these genes have shown G-protein-coupled γ-aminobutyric acid (GABA) receptor activity indicating a possible interaction between AtGLRs and GABA. Structurally, all of them were similar while differences were found regarding electrostatic surfaces as well as surface hydrophobicity. Results of this study provide a comprehensive reference regarding AtGLRs for further analysis regarding the structure, function, and evolution of the glutamate receptors in plants.

A strategy for designing "multi-prong" enzyme inhibitors by incorporating selective ligands to the liposomal surface.

We offer a novel strategy for designing "multi-prong" inhibitors of enzymes by incorporating selective ligands on the liposomal surface.

Ultrahigh resolution crystal structures of human carbonic anhydrases I and II complexed with "two-prong" inhibitors reveal the molecular basis of high affinity.

The atomic-resolution crystal structures of human carbonic anhydrases I and II complexed with "two-prong" inhibitors are reported. Each inhibitor contains a benzenesulfonamide prong and a cupric iminodiacetate (IDA-Cu(2+)) prong separated by linkers of different lengths and compositions. The ionized NH(-) group of each benzenesulfonamide coordinates to the active site Zn(2+) ion; the IDA-Cu(2+) prong of the tightest-binding inhibitor, BR30, binds to H64 of CAII and H200 of CAI. This work provides the first evidence verifying the structural basis of nanomolar affinity measured for two-prong inhibitors targeting the carbonic anhydrases.

Inhibition of matrix metalloproteinase-9 by "multi-prong" surface binding groups.

A novel strategy of blocking the active site accessibility of MMP-9 by "multi-prong" surface binding groups is described.

Solid-supported synthesis of polymerizable lanthanide-ion chelating lipids for protein detection.

Usually, lipids are synthesized employing solution-phase organic synthesis techniques. Though successful, the purifications can be difficult to accomplish due to the amphiphilic nature of the molecules. Herein, we demonstrate the advantages of a solid-phase approach for preparing a variety of metal-chelating lipids. A number of saturated and polymerizable metal-chelating lipids were prepared using this methodology. This approach requires one chromatographic purification after cleaving the lipids from the solid support. We also demonstrate that the resulting polymerized liposomes (containing Eu3+) possess the appropriate luminescence properties for the qualitative and quantitative determination of proteins.

Spacer-based selectivity in the binding of "two-prong" ligands to recombinant human carbonic anhydrase I.

Benzenesulfonamide and iminodiacetate (IDA)-conjugated Cu(2+) independently interact at the active site and a peripheral site of carbonic anhydrases, respectively [Banerjee, A. L., Swanson, M., Roy, B. C., Jia, X., Haldar, M. K., Mallik, S., and Srivastava, D. K. (2004) J. Am. Chem. Soc. 126, 10875-10883]. By attaching IDA-bound Cu(2+) to benzenesulfonamide via different chain length spacers, we synthesized two "two-prong" ligands, L1 and L2, in which the distances between Cu(2+) and NH(2) group of sulfonamide were 29 and 22 A, respectively. We compared the binding affinities of L1 and L2, vis-a-vis their parent compound, benzenesulfonamide, for recombinant human carbonic anhydrase I (hCA-I) by performing the fluorescence titration and steady-state kinetic experiments. The experimental data revealed that whereas the binding affinity of L1 for hCA-I was similar to that of benzenesulfonamide, the binding affinity of L2 was approximately 2 orders of magnitude higher, making L2 one of the most potent ligands or inhibitors of hCA-I. Since the enhanced binding or inhibitory potency of L2 is diminished (to the level of benzenesulfonamide) either in the presence of EDTA or upon treatment of the enzyme with diethyl pyrocarbonate, it is proposed that Cu(2+) of L2 interacts with one of the surface-exposed histidine residues of the enzyme. A cumulative account of the experimental data leads to the suggestion that the differential binding of L1 versus L2 to hCA-I is encoded in the chain length of the spacer moiety.

Evaluation of two lanthanide complexes for qualitative and quantitative analysis of target proteins via partial least squares analysis.

Two lanthanide complexes, namely 5-aminosalicylic acid ethylenediaminetetraacetate europium(III) (5As-EDTA-Eu3+) and 4-aminosalicylic acid ethylenediaminetetraacetate terbium(III), were evaluated for the analysis of carbonic anhydrase, human serum albumin (HSA), and gamma-globulin. Quantitative analysis is based on their luminescence enhancement upon protein binding and qualitative analysis on their lifetime capability to recognize the binding protein. Analytical figures of merit are presented for the three proteins. The limits of detection with 5As-EDTA-Eu3+ are at the parts per billion level. Partial least square regression analysis is used to determine HSA and gamma-globulin in binary mixtures without previous separation at the concentration ranges typically found in clinical tests of human blood serum.

Two-prong inhibitors for human carbonic anhydrase II.

The enzyme inhibitors are usually designed by taking into consideration the overall dimensions of the enzyme's active site pockets. This conventional approach often fails to produce desirable affinities of inhibitors for their cognate enzymes. To circumvent such constraints, we contemplated enhancing the binding affinities of inhibitors by attaching tether groups, which would interact with the surface exposed amino acid residues. This strategy has been tested for the inhibition of human carbonic anhydrase II. Benzenesulfonamide serves as a weak inhibitor for the enzyme, but when it is conjugated to iminodiacetate-Cu2+ (which interacts with the surface-exposed His residues) via a spacer group, its binding affinity is enhanced by about 2 orders of magnitude. This "two-prong" approach is expected to serve as a general strategy for converting weak inhibitors of enzymes into tight-binding inhibitors.

Protein surface-assisted enhancement in the binding affinity of an inhibitor for recombinant human carbonic anhydrase-II.

We elaborate on a novel strategy for enhancing the binding affinity of an active-site directed inhibitor by attaching a tether group, designed to interact with the surface-exposed histidine residue(s) of enzymes. In this approach, we have utilized the recombinant form of human carbonic anhydrase-II (hCA-II) as the enzyme source and benzenesulfonamide and its derivatives as inhibitors. The steady-state kinetic and the ligand binding data revealed that the attachment of iminodiacetate (IDA)-Cu(2+) to benzenesulfonamide (via a triethylene glycol spacer) enhanced its binding affinity for hCA-II by about 40-fold. No energetic contribution of either IDA or triethylene glycol spacer was found (at least in the ground state of the enzyme-inhibitor complex) when Cu(2+) was stripped off from the tether group-conjugated sulfonamide derivative. Arguments are presented that the overall strategy of enhancing the binding affinities of known inhibitors by attaching the IDA-Cu(2+) groups to interact with the surface-exposed histidine residues will find a general application in designing the isozyme-specific inhibitors as potential drugs.

An investigation on the analytical potential of polymerized liposomes bound to lanthanide ions for protein analysis.

We present a promising approach to protein sensing based on Eu3+ ions incorporated into polymerized liposomes. The sensitization of Eu3+ is accomplished with 5-aminosalicylic acid, which provides energy transfer for a stable reference signal and a wide wavelength excitation range free from protein interference. The lipophilic character of polymerized liposomes provides the appropriate platform for protein interaction with the lanthanide ion. Quantitative analysis is based on the linear relationship between the luminescence signal of Eu3+ and protein concentration. Because no spectral shift of the lanthanide luminescence is observed upon protein interaction, qualitative analysis is based on the luminescence lifetime of polymerized liposomes. This parameter, which changes significantly upon protein-liposome interaction, follows a well-behaved single-exponential decay that might be useful for protein identification.

Conjugation of poor inhibitors with surface binding groups: a strategy to improve inhibition.

Conjugation of surface binding groups with inhibitors for carbonic anhydrase leads to the conversion of weak inhibitors to strong inhibitors.

Synthesis of metal-chelating lipids to sensitize lanthanide ions.

Sensitization of lanthanide ions is important for lanthanide ion-based assays and sensing. To the best of our knowledge, there are very few reports of lanthanide ion sensitization after it is incorporated into the liposome surface. This paper describes the syntheses of several saturated and polymerizable metal-chelating lipids based on chelidamic acid. The lipids are synthesized either from (S)-ornithine or racemic 2,3-diaminopropanoic acid. These lipids as well as polymerized liposomes incorporating these lipids sensitize lanthanide ions. Liposomes from the lipid 18-Eu(3+) provided a probe that relies not only on the emission wavelengths of Eu(3+) but also on a reproducible lifetime that can be used for protein identification.

Synthesis of new, pyrene-containing, metal-chelating lipids and sensing of cupric ions.

The syntheses of several saturated, pyrene-containing, metal-chelating lipids are described. These lipids are capable of strongly binding to transition metal ions employing the metal-chelating headgroup. The excimer-to-monomer ratio of the pyrene groups changes with addition of cupric ions to the liposomes. Three other transition metal ions (Zn(2+), Ni(2+), and Hg(2+)) did not cause any appreciable changes in the excimer-to-monomer ratio. [reaction--see text]